Studies on dual-functioning of bio-absorbable scaffold materials and cells for the rehabilitation of oral tissues

生物可吸收支架材料与细胞双重功能用于口腔组织康复的研究

• 批准号: 12671880
• 负责人: TAIRA Masayuki
• 金额: $ 2.3万
• 依托单位: Iwate Medical University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2000
• 资助国家: 日本
• 起止时间: 2000 至 2002
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-12671880/
• 关键词: tissue engineering; bio-absorbable materials; cell culture; collagen gel; polylactide sponge; bone marrow stromal cells; osteogenic differentiation; gene analysis; コラーゲン; ゼラチン; 足場材料; ポリ乳酸; 遺伝子分析; チタン; アパタイト; アルジネート; 口腔組織; 組織再生

The findings obtained are described in the following four categories.(1) Studies on the preparation of bio-absorbable scaffold materials : We succeeded in producing inter-connected porous sponges by freeze-drying from natural materials (collagen and gelatin) and synthetic materials (alginate and poly(lactide-caprolacton)).(2) Studies on the cell culture : We investigated the effect of medium type (alfa-ME or DME), serum amount, dexamethasone induction and the growth factors (e.g. TGF-beta) on the proliferation and differentiation of rat bone marrow stromal cells and osteoblastic MC3T3-E1 cells. The former cells required the addition of vitamin C and the serum amount of more than 10% for the cell proliferation. The dexamethasone and the growth factors appeared to accelerate the osteogenic differentiation.(3) Studies on the duplex-functioning of the cells and scaffold materials : we successfully cultured the rat bone marrow stromal cells in the collagen gel frame-worked by poly (lactide- caprolacton) sponge. We examined the degree of osteogenic differentiation of rat bone marrow stromal cells cultured in the collagen gel. With the DME medium supplemented with vitamin C, the cells could differentiate into the pre-osteoblasts with their phenotype characterized by the appearance of mRNA for osteopontin. The cells in this stage appear to contribute to the rehabilitation of lost oral hard tissues in the future.(4) Studies on the tissue reaction against the scaffold materials : We implanted prepared scaffold materials (collagen gel, collagen sponge, poly (lactide-caprolacton) sponge, and gelatin hydro-gel) in the back tissue of SD rats, and evaluated the histo-pathological reaction against these scaffold materials. All scaffold materials were mildly absorbed by the host tissue without any strong pathological reaction.

所得结果分为以下四类。(1)生物可吸收支架材料制备研究：以天然材料（胶原蛋白和明胶）和合成材料（海藻酸盐和聚乳酸-己内酰胺）为原料，通过冷冻干燥法制备了互联多孔海绵。(2)细胞培养研究：研究培养基类型（α - me或DME）、血清量、地塞米松诱导及生长因子（如tgf - β）对大鼠骨髓基质细胞和成骨细胞MC3T3-E1增殖分化的影响。前一种细胞需要添加维生素C和10%以上的血清量才能使细胞增殖。地塞米松和生长因子有加速成骨分化的作用。(3)细胞与支架材料的双重功能研究：我们成功地将大鼠骨髓基质细胞培养在聚乳酸-己内酰胺海绵的胶原凝胶框架中。我们观察了胶原凝胶培养大鼠骨髓基质细胞的成骨分化程度。在添加维生素C的DME培养基中，细胞可分化为成骨前细胞，其表型以骨桥蛋白mRNA的出现为特征。这一阶段的细胞似乎有助于未来口腔硬组织的恢复。(4)组织对支架材料的反应研究：将制备好的支架材料（胶原凝胶、胶原海绵、聚乳酸-己内酰胺海绵、明胶水凝胶）植入SD大鼠背部组织，评价其对支架材料的组织病理反应。所有支架材料均被宿主组织轻度吸收，无强烈病理反应。

项目成果

TAIRA, M., ARAKI, Y. and TAKAHASHI, J.: "Scaffold consisting of poly(lactide-caprolacton) sponge, collagen gel and bone marrow stromal cells for tissue engineering"J.Mater.Sci.Lett.. 20(19). 470-477 (2001)

TAIRA, M., ARAKI, Y. 和 TAKAHASHI, J.：“用于组织工程的由聚（丙交酯-己内酯）海绵、胶原凝胶和骨髓基质细胞组成的支架”J.Mater.Sci.Lett.. 20(19)

M.Taira,M.S.Toguchi,Y.Hamada,M.Okazaki,J.Takahashi,R.Itoh,S.Toyosawa and N.Ijyuin: "Studies on cytotoxicity of nickel ions using C3H10T1/2 fibroblast cells"Journal of Oral Rehabilitation. 27・6. 1068-1072 (2000)

M.Taira、M.S.Toguchi、Y.Hamada、M.Okazaki、J.Takahashi、R.Itoh、S.Toyosawa 和 N.Ijyuin：“使用 C3H10T1/2 成纤维细胞研究镍离子的细胞毒性”口腔康复杂志。 27・6。1068-1072（2000）

M.Taira,M.S.Toguchi,Y.Hamada,J.Takahashi,R.Itoh,S.Toyosawa,N.Ijyuin and M.Okazaki: "Studies on cytotoxic effect of nickel ions on three cultured fibroblasts"Journal of Materials Science : Materials in Medicine. 12(印刷中). (2001)

M.Taira、M.S.Toguchi、Y.Hamada、J.Takahashi、R.Itoh、S.Toyosawa、N.Ijyuin 和 M.Okazaki：“镍离子对三种培养成纤维细胞的细胞毒性作用的研究”材料科学杂志：材料12（正在出版）（2001）。

Taira, M., Nakao, H., Takahashi, J., Araki, Y.: "Effects of two vitamins, two growth factors and dexamethasone on the proliferation of rat bone marrow stromal cells and osteoblastic MC3T3-E1 cells"J.Oral Rehabilitation. 30(印刷中). (2003)

Taira, M.、Nakao, H.、Takahashi, J.、Araki, Y.：“两种维生素、两种生长因子和地塞米松对大鼠骨髓基质细胞和成骨细胞 MC3T3-E1 细胞增殖的影响”J.Oral康复。30（正在出版）（2003）。

TAIRA, M., NAKAO, H., TAKAHASHI, J. and ARAKI, Y.: "Cellular reactions to polylactide-based sponge and collagen gel in subcutaneous tissue"J.Oral Rehabil.. 30(1). 106-109 (2003)

TAIRA, M.、NAKAO, H.、TAKAHASHI, J. 和 ARAKI, Y.：“皮下组织中聚丙交酯海绵和胶原凝胶的细胞反应”J.Oral Rehabil.. 30(1)。